CN109576323A - The sequential feeding method of the enzymatic hydrolysis of pretreated base-material is gradually prolongedly added at any time - Google Patents
The sequential feeding method of the enzymatic hydrolysis of pretreated base-material is gradually prolongedly added at any time Download PDFInfo
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Abstract
The present invention relates to the sequential feeding methods of enzymatic hydrolysis, wherein under stiring, contact pretreated lignocellulosic base-material with water and enzyme in the reactor, the method is characterized in that adding pretreated lignocellulosic base-material to reactor sequence and is carried out in a manner of gradually extended at any time, to obtain scheduled final dry matter content.
Description
Technical field
The present invention relates to such as undefined with sequence supplying mode by pretreated lignocellulosic base-material
(substrate) the enzymatic hydrolysis method carried out, so that the cellulose is converted to glucose.Then, the glucose can be with
It is used in various further steps, such as in the fermentation step of production alcohol, or for producing chemical intermediate.
Background technique
The economically viable method of exploitation modification lignocellulose biomass is currently " hot topic ".Fossil resource
New way that is constantly short and already leading to researching and producing bio-fuel and chemical intermediate with the competition of food supply.
Since nineteen seventies, the conversion of the lignocellulose biomass after fraction polysaccharide hydrolyzes saccharogenesis
Just have become the theme much studied.
Lignocellulose biomass is characterized in that by three kinds of principal polymerics: cellulose, hemicellulose and lignin structure
At labyrinth, the ratio of cellulose, hemicellulose and lignin changes with the Change of types of lignocellulose biomass.
A kind of typical but unrestricted composition is as follows: cellulose is with 35% to 50% amount presence, hemicellulose, be substantially by
The polysaccharide that pentose and hexose are constituted, exists with 20% to 30% amount and lignin exists with the amount of 15 weight % to 25 weight %.?
Confirm that the degradation of biomass is highly difficult, because the polysaccharide (cellulose and hemicellulose) of plant walls is closely combined with lignin,
This provides hardness for wall.
In these three polymer, cellulose is main sugar source, because it is made of glucose；Glucose is easy to change
Matter.
In general, including multiple steps by the method for bio-chemical pathway modifying bio matter.First step be collect and it is defeated
Send lignocellulose biomass to biomass transformation center.Second step is the pretreatment or prehydrolysis of biomass, makes fibre
Therefore the accessible enzyme of dimension element simultaneously can generate pretreated lignocellulosic base-material.Third step, enzymatic hydrolysis, refer to due to
Use the solution (also referred to as enzymatic mixture) of the cellulolytic enzyme and hydrolysis of hemicellulose enzyme generated by microorganism, cellulose
It is converted to glucose.Then glucose can generally pass through saccharomyces cerevisiae (yeast in the four steps of fermentationSaccharomyces cerevisiae) it is modified to intermediate product, such as ethyl alcohol, or pass through clostridium acetobutylicum yeast (yeastClostridium acetobutylicum) modified by fermentation as acetone, butane, ethyl alcohol (ABE) mixture.Then, the 5th
Step distillation, which refers to, can be concentrated resulting molecule.Glucose can also modify as bio-fuel (hydrogen, methane).
Therefore, a committed step is enzymatic hydrolysis.In enzymatic hydrolysis step, the pretreated lignocellulosic
Base-material must be mixed with the liquid solution comprising cellulolytic enzyme and hydrolysis of hemicellulose enzyme.Since purpose is to obtain high concentration
Sugar, thus enzymatic hydrolysis step must with the pretreated lignocellulosic base-material (i.e. with high dry content) of high concentration into
Row.It has been evaluated that, sugared concentration (its dry for corresponding approximately to 15 weight % of minimum of 8 weight % is generated during enzymatic hydrolysis
Matter content) when, the method be it is economically feasible (McIntosh, S., Zhang, Z., Palmer, J., Wong,
H., Doherty, W.O.S., Vancov, T., 2016. Pilot-scale cellulosic ethanol
production using eucalyptus biomass pre-treated by dilute acid and steam
explosion. Biofuels, bioproducts and biorefining 10 (4), 346-358).In high dry matter
Operating under content also reduces reactor volume, and therefore this method finance and cost of energy reduce (Larsen, J.,
Ostergaard Petersen, M., Thirup, L., Wen Li, H., Krogh Iversen, F., 2008. The
IBUS process of lignocellulosic bioethanol close to a commercial reality.
Chem. Eng. Technol. 31, 765-722)。
However, being able to verify that when dry matter content is high, pretreated lignocellulosic base-material and containing cellulose are hydrolyzed
The close mixing of the liquid solution of enzyme and hydrolysis of hemicellulose enzyme is difficult.In fact, being started with high dry content
Enzymatic hydrolysis generates the particular problem of mixing and homogenizing.Reaction medium is very sticky, needs special stirring, this is more mixed than in reaction
The stirring needed at the end of closing hydrolysis when object has become more fluidization is much more complex.
In general, enzymatic hydrolysis can carry out in discontinuous or flow reactor.In discontinuous or batch process
In, by whole components, the substance including controlling pH is put into reactor when hydrolyzing and starting.During Hydrolyze method, not to anti-
It answers device to add any substance or does not take out any substance from reactor.In a continuous process, there are both inlet flow and output stream,
But reaction volume is kept constant.
In another configuration of the method, it is also referred to as sequential feeding or batch feed mode method, is carried out in method
Period does not take out any substance from reactor, but gradually adds base-material to reactor in a sequential manner without taking out during hydrolysis
Any hydrolysate.It has been found that and is fed using such base-material, such as products collection efficiency as caused by base-material can be overcome
Inhibition influence.With reaction progress and when reacting progress, mixture gradually becomes liquid and can add fresh base-material
To improve dry matter content.Then it can reach high base concentration and be advantageously 17 to 30 weight % dry matters.
From the prior art (Mondebach, A.A., Nokel, S.E., 2013. Enzymatic hydrolysis
of biomass at high-solids loadings – A Review. Biomass and Bioenergy 56, 526-
544) the known enzymatic hydrolysis method using continuous feed.
Similarly, patent application US2010/330638A describes the confession in the batch feed mode of enzymatic hydrolysis step
Material shows that test can measure the amount for the biomass that can be added to each batch.Therefore, it when changing binder types every time, needs
It to be tested before enzymatic hydrolysis step.
Application WO2016/062646 describes a kind of method for preparing sugar and/or tunning by ligno-cellulosic materials,
It includes multiple enzymatic hydrolysis steps, and the first step is batch feed mode.
Patent application US 2010/0255554 describes a kind of with batch feed mode hydrolysis of lignocellulose biomass
Method, wherein fed by controlling reactor volume and/or the pretreated lignocellulose biomass of addition (and optionally enzyme
Addition) frequency and the sugared volume and/or concentration that generates in the reactor adjust the functional parameter of this method.Specifically, institute
It states pretreated lignocellulose biomass charging and reactor is always added to identical frequency.
Applicant has developed a kind of improved enzymatic hydrolysis methods using sequential feeding (batch feed), can
The glucose for obtaining high yield reduces the energy consumption and incorporation time of this method simultaneously.
More particularly it relates to a kind of sequential feeding method of enzymatic hydrolysis, wherein under stiring, making at least one
Pretreated lignocellulosic base-material is contacted with water and enzyme in the reactor, and the method is characterized in that pretreated wood
Matter cellulose base-material sequence is added to reactor and is carried out in a manner of gradually extended at any time, to obtain scheduled final dry
Matter content.
When by pretreated lignocellulosic base-material and preferred enzyme, with gradually extended sequential feeding is (in batches at any time
Charging) in injection enzymatic hydrolysis method when, compared to wherein with the batch feed mode of constant time interval addition base-material, seeing
Observe the raising of glucose yield and the reduction of energy consumption.
In addition, when in the same way add enzyme when, i.e., sequentially and at any time extend, and preferably with base-material simultaneously, note
It anticipates to the synergistic effect to glucose yield, is had not seen when adding whole enzymes when hydrolyzing and starting.
It is one advantage of the present invention that it provides a kind of enzymatic hydrolysis method for improving glucose yield.
Another advantage of the present invention provides a kind of usual due to batch feed technology (but it extends at any time) for it
The less enzymatic hydrolysis method for seeing mixing and viscosity problem (if present).In fact, gradually due to dry matter content
Increase, each base-material is added, is easy to be mixed, so that on the one hand being reduced and/or being used by the energy of blender consumption
Less complicated blender (such as dihedral vane formula impeller or marine impellers).
In addition, of the invention one further advantage is that provide a kind of enzymatic hydrolysis side that blender rotation speed is low
Method, for keeping critically important (Mhlongo SI, Haan R, Viljoen-Bloom M, the Zyl WH. of enzymatic activity
Lignocellulosic hydrolysate inhibitors selectively inhibit/deactivate
cellulase performance (2015). Enzyme and Microbial Technology, 81: 16–22)。
In addition, when handling (coprocessing) a variety of different types of base-materials simultaneously in same reactor, it is according to the present invention
Method is appropriate.
Another advantage of the invention for its provide one kind can monitor reaction medium variation and to its it is simple adapt to without
Want the complicated enzymatic hydrolysis method measured.
According to a variant, the final content of dry matter is higher than 12 weight %, and preferably 18 to 24 weight %.
According to a variant, enzyme is added to reactor sequentially and is gradually prolongedly carried out at any time.
According to a variant, pretreated lignocellulosic base-material and enzyme are added to reactor simultaneously.
According to a variant, for adding every time, pretreated lignocellulosic base-material adds in the same amount.
According to a variant, for adding every time, enzyme adds in the same amount.
According to a variant, reactor includes blender, and the ratio D:T of stirrer diameter and reactor diameter is 0.3
To 0.75.
According to a variant, blender is dihedral vane formula impeller or marine impellers.
According to a variant, enzyme is with the contact of 0.1 to 60 mg enzyme/g cellulose concentration.
According to a variant, temperature of the method at 40 to 60 DEG C is carried out under 4 to 6 pH and atmospheric pressure.
According to a variant, or mixtures thereof various pretreated lignocellulosic base-materials itself are used.
Fermentation step according to a variant, in the presence of being carried out after the method in the microorganism of production alcohol.
According to another variant, the method is according to the method for being saccharified and fermenting while referred to as SSF method in production alcohol
It is carried out in the presence of microorganism.
Summary of the invention
By wooden (fallen leaves property and resinite (resinous)) unprocessed or processed agricultural by product such as rice
Grass, plant fiber, forestry crop, production alcohol-, sugared and cereal plant residue, the residue from paper mill, marine biomass
(such as macro cellulosic residue) or ligno-cellulosic materials conversion product obtain pretreated lignocellulosic biomass
Matter.
Used lignocellulose biomass is preferably timber, wheat straw, wood pulp, awns genus plant, straw or corn stover.
Method according to the invention it is possible to use or mixtures thereof various types of lignocellulose biomass itself.
Lignocellulosic base-material used in the method for the present invention is to allow the physics by changing ligno-cellulosic materials
The result of preprocessing biomass under conditions of lignocellulosic is reconstructed with physicochemical property.Those skilled in the art can be used
The pretreatment for any kind of lignocellulose biomass known carries out pre-treatment step.It can also carry out preconditioned step, example
It such as include crushing or removing stone.The pre-treatment step may include heat, chemistry, mechanical and/or enzymatic treatment or the group of these processing
It closes.
According to a preferred variant, the pre-treatment step is selected from pretreatment in acid condition, such as in acidity
Under the conditions of acid cooking or steam blasting, the pretreatment in alkaline medium, for example, by using the pretreatment (side Kraft of vulcanized sodium
Method), ammonia circulation percolation (ARP) or ammonia fiber blasting procedure (AFEX), oxidation pre-treatment, for example, by using ozone, hydrogen peroxide, oxygen
The pretreatment of gas or peracetic acid, the pretreatment for not adding chemical reagent, for example, do not add acid steam blasting or by using pole
The pretreatment of hot water washing or organic solvent method.
The pretreatment that the pre-treatment step is carried out advantageously by steam blasting in acid condition.150 to 250
DEG C optimal conditions under carry out a few minutes.
The present invention relates to a kind of sequential feeding methods of enzymatic hydrolysis, wherein under stiring, making pretreated wooden fibre
It ties up plain base-material to contact with water and enzyme in the reactor, the method is characterized in that adding pretreated wood to reactor sequence
Matter cellulose base-material is carried out in a manner of gradually extended at any time, to obtain scheduled final dry matter content.
The scheduled final dry matter content is preferably higher than 12 weight %, preferably 15 to 30 weight %, and most preferably 18 to 24
Weight %.In remaining content, the concentration of pretreated lignocellulosic base-material is expressed as the weight percent of dry matter.
According to standard ASTM E1756-08 (2015) " Standard Test Method for Determination of Total
Solids in Biomass " measures dry matter content.
Dry matter content one when adding pretreated lignocellulosic base-material for the first time, when method for hydrolysis starts
As be lower than 10 weight %, preferably shorter than 8 weight %, especially preferably less than 6 weight %.
The frequency for adding pretreated lignocellulosic base-material gradually extends.Therefore, what is " gradually extended at any time " adds
Add and refers to the reduced frequency of use or be in other words added using the increased period.Therefore, n-th and it is added (n+1)th time
Between time passage less than (n+1)th time addition the n-th+2 times addition between time passage, and so on.Such as the first time and
Time passage between second of addition base-material is less than the time passage between third time addition second, and so on.Example
Such as, addition can occur after 1 hour for the first time, and second addition can occur after 3 hours, and third time addition can be with
Occur after 6 hours, the 4th addition can occur after 13 hours and the 5th addition can be sent out at 24 hours later
It is raw.
In order to reach scheduled final dry matter content, it is however generally that, with gradually extension system, carry out at least base-material three times
Addition, preferably at least No. four base-material additions, more preferably at least No. five base-material additions.
The amount added when adding pretreated lignocellulosic base-material typicallys represent most 5 weight %, preferably 2 to 5 weights
Measure the increase of the dry matter content of % and more preferable 2 to 3 weight %.The scale example such as 3 weight %'s of the base-material added in addition
Dry matter content.
According to a variant, the pretreated lignocellulosic base-material is added in each addition with equal amount.
According to a preferred variant, sequentially and at any time gradually prolongedly carried out to reactor addition enzyme.It has seen
It observes and to maintain enzymatic activity at any time with extension, batch feed mode addition enzyme, this is added entirely with when enzymatic hydrolysis starts
Portion's enzyme is different.
The pretreated lignocellulosic base-material and enzyme simultaneously or staggeredly can be added to reactor, while at any time
Gradually prolongedly continuation sequence adds each component.The pretreated lignocellulosic base-material and enzyme are preferably added to anti-simultaneously
Answer device.
According to a variant, enzyme is added in each addition with equal amount.
The amount being added in addition is generally 0.1 to 60 mg enzyme/g cellulose, preferably 5 to 40 mg enzymes/g cellulose, most
It is preferred that 10 to 30 mg enzymes/g cellulose.
The enzymatic hydrolysis generally carries out under 4 to 6, preferably 4.5 to 5.8, more preferable 4.8 to 5.5 pH.It generally exists
It is carried out at a temperature of 40 to 60 DEG C, preferably 45 to 55 DEG C.It is advantageously carried out at atmosheric pressure.
The enzymatic hydrolysis is carried out by the enzyme generated by microorganism.Added enzyme solutions include by cellulose decomposition at
The enzyme of sugar.Microorganism, such as the fungi or anaerobic bacteria of trichoderma, aspergillus, Penicillium or Schizophyllum, such as fusobacterium
Anaerobic bacteria, generate these enzymes, the cellulase largely hydrolyzed particularly with suitable cellulose.With highly preferred side
Formula, the cellulolytic enzyme of step d) by microorganism trichoderma reesei (Trichoderma reesei) generate.
According to the present invention, time of contact when enzymatic hydrolysis be 5 to 200 hours, preferably 2 to 100 hours, most preferably 1 to
50 hours.
Method according to the present invention can be and measuring the value of a rheological properties of reaction medium at any time
Monitoring, the rheological properties be advantageously selected from reaction medium viscosity, stirring system axis torque and motor consumed by electric work
Rate.Electrical power consumed by motor has symbol PElectricity。
During the method for the present invention, i.e., during liquefaction, the torque and motor of the viscosity of reaction medium, stirring system axis
Consumed electrical power is the rheological properties useful to lignocellulosic base-material caused by monitoring from many aspects.It is true
On, these characteristics of viscosity, torque and power are inter-related.Electrical power P consumed by motorElectricityWith driving blender axis
Mechanical output PIt is mechanicalIt is associated.
Electrical power consumed by motor is the parameter of general measure and monitoring in pilot scale or the equipment of industrial product.
Following formula defines the relationship between various parameters:
PIt is mechanical = f (PElectricity), f is the motor design particularities illustrated by motor construction side.
PIt is mechanical=2 π N*C, in which:
N is the mixing speed in terms of revolutions per second,
C is the torque in terms of N.m,
And PIt is mechanicalFor with the power of wattmeter.
During churning, following relationship is applicable in:
PIt is mechanical = ρNpN3D5
ρ is with kg.m-3The density of the reaction medium of meter,
D is the diameter of the blender in terms of m,
NpFor the characteristic of blender, the geometry and flow pattern of slot are depended on.
During laminar flow flow pattern, following relationship is applicable in:
Np=A/Re, therefore PIt is mechanical = ρAN3D5/Re
Wherein A is stirring system constant, and Re is Reynolds number, wherein Re=ρ ND2/,
Average dynamic viscosity for the reaction medium measured in terms of pascal second (Pa.s), wherein = PIt is mechanical/(AN2D3) =
2πC/(AD3N) 。
Although the torque of viscosity and stirring system axis is the measurement to be easy to get on a small scale, electric work consumed by motor
Rate PElectricityFor the magnitude for being easiest to measurement at industrial scale.
In a manner of highly preferred, method according to the present invention be characterized in that electrical power consumed by motor with
Time measures.
Method according to the present invention advantageously carries out in the reactor, and the reactor preferably has cylinder, high
Degree/diameter ratio is advantageously 1 to 3.
Since gradually extended batch feed mode, the influence of reaction medium medium viscosity are less significant.Routinely, selected
Blender allow for processing laminar flow.Then, wide blender, or scratch reactor wall even under medium rotational specd and apply
Those of blending and kneading effect is added to be necessary.In the method according to the invention, can be used dihedral vane formula impeller or
The better simply blender of marine impellers type.
Specifically, the blender with small diameter can be used in the method according to the invention.According to a variant,
Stirrer diameter/reactor diameter ratio D/T is advantageously 0.3 to 0.75, preferably 0.4 to 0.65.
Similarly, rotation speed can be lower than conventional system.Rotation speed is generally below 100 rpm(revolutions per minutes),
Preferably shorter than 80 rpm.
According to a preferred embodiment, can be carried out after enzymatic hydrolysis method according to the present invention by producing alcohol
The alcohol method fermentation step that carries out of microorganism to generate the fermentation effluent of alcohol-containing.
The enzymatic hydrolysis and the fermentation of alcohol method can also carry out simultaneously.It is synchronous glycosylation and fermentation or SSF herein
The case where method.Enzymatic hydrolysis can also be carried out according to other arrangements well known by persons skilled in the art and alcohol method is fermented, such as
Synchronous glycosylation and fermentation process (PSSF) are carried out after premashing or mix hydrolysis and fermentation process (HHF).
Alcohol can be fermented by the sugar that enzymatic hydrolysis obtains, such as ethyl alcohol, 1,3-PD, isopropanol, n-butyl alcohol, different
Or mixtures thereof butanol or 1,4- butanediol itself.The alcohol method fermentation is preferred to generate ethyl alcohol.
Ensure that alcohol method is fermented by the microorganism of yeast or other production alcohol.In the meaning of the present invention, term " alcohol method hair
Ferment " refers to method of the sugar only by microbial fermentation at one or more alcohol.It is raw used in the alcohol method fermentation step of hexose
The microorganism for producing alcohol is preferably selected from yeast and bacterium, can be transgenosis.
When the microorganism for producing alcohol is yeast, saccharomyces cerevisiae is most effective.Also it can choose yeast such as grain wine to split
Grow yeast (Schizosaccharomyces pombe) or saccharomyces uvarum (Saccharomyces uvarum) or saccharomyces diastaticus
(diastaticus).More thermophilic yeast, such as Kluyveromyces fragilis (Kluyveromyces fragilis) (now logical
Frequently referred toK. marxianus) be also advantageous, especially when enzymatic hydrolysis and alcohol method ferment while carrying out (SSF method).
Genetically modified organism, such as saccharomyces cerevisiae type yeast such as 3400 (Ohgren et al., J. of TMB can also be used
of Biotech 126, 488-498, 2006)。
When produce alcohol microorganism be bacterium when, preferably for example zymomonas mobilis (Zymomonas mobilis), the effective way of the assimilation of production ethyl alcohol or the anaerobic bacteria of fusobacterium are provided, such as pure and mild molten for producing
The mixture of agent such as acetone-butanol-ethanol (ABE) or isopropanol-butanol-ethyl alcohol (IBE) clostridium acetobutylicum, or be used for
Produce the Escherichia coli of isobutanol.
The fermentation of alcohol method preferably 30 DEG C to 40 DEG C at a temperature of carry out at 3 to 6.5 pH.
Yeast, preferably saccharomyces cerevisiae are used highly preferred microorganism.They have bigger robustness and peace
Quan Xing, and do not need the aseptic condition of operation the method and equipment.
The yeast of saccharomyces being capable of only zymohexose (substantially glucose and mannose).These yeast are in an optimized way
Hexose is modified to ethyl alcohol and good conversion ratio can be obtained.
When carrying out enzymatic hydrolysis and alcohol method fermentation (SSF method) in identical operation, temperature is preferably 30 to 45 DEG C,
PH is 4 to 6, to stimulate yeast performance.
Operation embodiment hereafter is intended to illustrate the present invention.
Specific embodiment
By have the warp of high dry matter (DM) content to locate in advance with gradually extension system sequential feeding (batch feed)
The enzymatic hydrolysis method of the wheat straw of reason.It is different from conventional batch feed strategies (it is added at regular intervals), it is this new
Strategy is constituted by sequentially increasing dry matter content in a manner of gradually extending at any time and add.Fig. 1 a and 2a are shown according to routine
Batch feed technology addition, and Fig. 1 b and 2b are shown according to the gradually addition of extended batch feed technology at any time.Fig. 2
The details of Fig. 1 is exaggerated across time coordinate.
The whole water contents (1.3 kg) for being enough to realize 5 weight % dry substance concentrations are loaded into reactor when testing and starting
With the pretreated wheat straw base-material (250 g) of first time addition.Then, 170 g are carried out after 1,3,6,13 and 24 hour
Five equivalent additions of lignocellulosic matrix are to realize the dry matter (Fig. 1 b and 2b) of 20 weight %.In the same way into
The addition of row enzyme.Using this strategy, without departing from critical viscosity value, the gradually liquefaction of base granule is possible
, because this does not allow being sufficiently mixed using dihedral vane formula or marine impellers.
80 to 85% glucose yield is allowed for according to the method for the present invention, wherein the enzymatic hydrolysis through 48 h has
The low-energy-consumption of 35-40 kJ.Specifically, by the method for the invention through 48 h realize energy consumption and using other in batches
The energy consumption that feed strategies are only realized in 5 h is identical.In fact, when cellulose is high living by existing in cellulase conversion
When property, the exponential increase of glucose is observed within 24 hours at first of test.Later, increase slack-off: within preceding several hours,
The every kJ consumed in mixture the glucose for recording 50 to 90 g is increased；It after 24 hours, is 10-15 g/kJ, and
After 48 h, 10 g/kJ is fallen below.
Similarly, the rotation speed of hybrid system is slow (about 80 rpm).The enzyme is with stable by weak force
Molecular structure protein.This weak steady refers to that various parameters influence protein.Mechanical stress is that can reduce enzymatic activity
Factor.
Gradually the strategy of extended sequence addition can also be suitable for enzyme at any time.Fig. 3 shows the addition when testing beginning
The enzyme (ZE, Fig. 3) of whole amount and adjoint sequence add base-material and sequentially add the difference between (GE, Fig. 3).
If Fig. 3 shows the enzyme (52.8 g) for feeding whole amount when enzymatic hydrolysis starts, glucose is quickly
Increase.However, it means that generation glucose is too fast in reaction medium, inhibits enzyme and therefore inhibits glucose generation.Phase
Than under, gradually add enzyme (for adding every time, 8.8 g, at any time gradually extend) make inhibit enzyme substituted and
The glucose (Fig. 3) of high yield is obtained at the end of this method.
Claims (14)
1. the sequential feeding method of enzymatic hydrolysis, wherein under stiring, making at least one pretreated lignocellulosic base-material
It is contacted in the reactor with water and enzyme, the method is characterized in that adding pretreated lignocellulosic to reactor sequence
Base-material is carried out in a manner of gradually extended at any time, to obtain scheduled final dry matter content.
2. according to the method described in claim 1, wherein the final dry matter content is higher than 12 weight %.
3. method according to claim 1 or 2, wherein the final dry matter content is 18 to 24 weight %.
4. method according to any one of claims 1 to 3, wherein sequentially and at any time gradually prolonging to reactor addition enzyme
It carries out longly.
5. method according to any one of claims 1 to 4, wherein the pretreated lignocellulosic base-material and enzyme are same
When be added to reactor.
6. method according to any one of claims 1 to 5, wherein for adding every time, the pretreated wooden fibre
Plain base-material is tieed up to add in the same amount.
7. method according to any one of claims 1 to 6, wherein enzyme adds in the same amount for adding every time.
8. method according to any one of claims 1 to 7, wherein the reactor includes blender, and blender
The ratio D:T of the diameter of diameter and reactor is 0.3 to 0.75.
9. method according to any one of claims 1 to 8, wherein the blender is dihedral vane formula impeller or leaf peculiar to vessel
Wheel.
10. method according to any one of claims 1 to 9, wherein the enzyme is dense with 0.1 to 60 mg enzyme/g cellulose
Degree contact.
11. the method according to any of claims 1 to 10, wherein the method 40 to 60 °C at a temperature of, 4 to
It carries out under 6 pH and at atmosheric pressure.
12. according to claim 1 to 11 described in any item methods, wherein using various pretreated lignocellulosic base-materials
Or mixtures thereof itself.
13. wherein carrying out the microorganism in production alcohol after the method to 12 described in any item methods according to claim 1
In the presence of fermentation step.
14. according to claim 1 to 12 described in any item methods, wherein the method is according to the method for synchronous glycosylation and fermentation
It is carried out in the presence of producing the microorganism of alcohol.
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| FR1759033A FR3071510B1 (en) | 2017-09-28 | 2017-09-28 | ENZYMATIC HYDROLYSIS PROCESS IN SEQUENTIAL FEEDING WITH ADDITIONS OF PRE-TREATED SUBSTRATE IN MORE AND MORE SPACES OVER TIME |
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| CN104651429B (en) | 2015-02-03 | 2018-05-18 | 湖南农业大学 | A kind of method for irradiating pretreatment and the batch feeding realization high concentration of substrate enzyme hydrolysis of lignocellulosic |
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| CARDONA MJ等: "A process for energy-efficient high-solids fed-batch enzymatic liquefaction of cellulosic biomass", 《BIORESOURCE TECHNOLOGY》 * |
| FAN ZL等: "Conversion of paper sludge to ethanol. I: Impact of feeding frequency and mixing energy characterization", 《BIOPROCESS BIOSYST ENG》 * |
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| Publication number | Publication date |
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| ES2852425T5 (en) | 2024-06-20 |
| EP3461902B1 (en) | 2020-12-02 |
| CN109576323B (en) | 2023-07-04 |
| EP3461902B2 (en) | 2024-01-03 |
| EP3461902A1 (en) | 2019-04-03 |
| DK3461902T4 (en) | 2024-03-25 |
| FR3071510A1 (en) | 2019-03-29 |
| CA3018686A1 (en) | 2019-03-28 |
| ES2852425T3 (en) | 2021-09-13 |
| FR3071510B1 (en) | 2021-08-27 |
| US10865426B2 (en) | 2020-12-15 |
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| HRP20210325T1 (en) | 2021-04-02 |
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| BR102018069236A2 (en) | 2019-04-16 |
| US20190093133A1 (en) | 2019-03-28 |
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